Journal: ACS Chemical Neuroscience

Article Title: Interplay of Neuroinflammation and Gut Microbiota Dysbiosis in Alzheimer’s Disease Using Diffusion Kurtosis Imaging Biomarker in 3 × Tg-AD Mouse Models

doi: 10.1021/acschemneuro.5c00063

Figure Lengend Snippet: Comprehensive experimental design for assessing AD pathology in mice. The timeline illustrates the multistep workflow for analyzing behavioral, imaging, microbiota, cytokine, and neuroinflammatory changes in 6-MO and 12-MO WT and AD mice ( N = 5 per group). On day 1, cognitive performance was evaluated using the T-maze task to assess working memory. On day 2, MRI/DKI was conducted to analyze microstructural brain changes. On day 3, fecal samples were processed with NGS to evaluate gut microbiota composition and diversity. On day 4, ELISA was used to quantify systemic pro-inflammatory cytokines, including IL-1β, IL-6, TNFα, and IFN-γ. Meanwhile, the serum samples were quantified for SCFA levels to assess dysregulated gut metabolism. On day 5, IHC was performed to assess microglial (Iba1) and astrocytic (GFAP) activation in key brain regions such as the mPFC, HIPP, and STR. This comprehensive workflow integrates multiple methodologies to investigate age- and disease-specific changes in AD pathology.

Article Snippet: Cytokine levels (IL-1β, IL-6, IFN-γ, and TNFα) were quantified using a multiplex ELISA kit (MEK1016; Boster, Pleasanton, CA, USA).

Techniques: Imaging, Enzyme-linked Immunosorbent Assay, Activation Assay

Journal: BMC Veterinary Research

Article Title: Quick and improved immune responses to inactivated H9N2 avian influenza vaccine by purified active fraction of Albizia julibrissin saponins

doi: 10.1186/s12917-020-02648-1

Figure Lengend Snippet: Effect of AJSAF on H9–Ag-specific IgG and its isotype antibody titers in the immunized mice. Mice were s.c. injected with 0.2 ml of IH9V (10 7 TCID 50 /0.1 ml) alone or containing Quil A or AJSAF, or 0.2 ml of CH9V (10 7 TCID 50 /0.1 ml) on days 1 and 15. Sera were collected 14 days after the boosting immunization, and serum H9–Ag-specific IgG, IgG1, IgG2a, and IgG2b antibody titers were measured by ELISA. The values are presented as means ± SD ( n = 6). Significant differences with IH9V alone group were designated as * P < 0.05, † P < 0.01, and ‡ P < 0.001; those with CH9V group as § P < 0.05, # P < 0.01, and ¶ P < 0.001

Article Snippet: Assoc., Birmingham, AL, USA; rabbit anti-chicken IgY horseradish peroxidase conjugate was from Promega Corporation, Madison, WI, USA; mouse cytokine detecting ELISA kits were from Wuhan Boster Bio-Tech.

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: BMC Veterinary Research

Article Title: Quick and improved immune responses to inactivated H9N2 avian influenza vaccine by purified active fraction of Albizia julibrissin saponins

doi: 10.1186/s12917-020-02648-1

Figure Lengend Snippet: Effect of AJSAF on haemagglutination inhibition (HI) and H9–Ag-specific IgY antibody levels in the immunized chicken. The chickens were prime- and boost-immunized with 0.4 ml of IH9V (10 7 TCID 50 /0.1 ml) alone or containing AJSAF, or 0.4 ml of CH9V (10 7 TCID 50 /0.1 ml) at 3-week interval. Sera were collected on designated days post-immunization, and the serum HI antibody titers ( a ) and H9–Ag-specific IgY antibody ( b ) levels were measured by HI assay and ELISA, respectively. The values are presented as means ± SD ( n = 30). Significant differences with IH9V alone group were designated as * P < 0.05, † P < 0.01, and ‡ P < 0.001

Article Snippet: Assoc., Birmingham, AL, USA; rabbit anti-chicken IgY horseradish peroxidase conjugate was from Promega Corporation, Madison, WI, USA; mouse cytokine detecting ELISA kits were from Wuhan Boster Bio-Tech.

Techniques: Inhibition, HI Assay, Enzyme-linked Immunosorbent Assay

Journal: BMC Veterinary Research

Article Title: Quick and improved immune responses to inactivated H9N2 avian influenza vaccine by purified active fraction of Albizia julibrissin saponins

doi: 10.1186/s12917-020-02648-1

Figure Lengend Snippet: Effect of AJSAF on cytokine secretion from splenocytes in the immunized mice. Splenocytes were incubated with Con A (48 h) or H9–Ag (72 h), and the supernatants were collected for detecting IL-2, IFN-γ, and IL-10 levels using ELISA kits. The values are presented as means ± SD ( n = 6). Significant differences with IH9V alone group were designated as * P < 0.05, † P < 0.01, and ‡ P < 0.001; those with CH9V group as # P < 0.01 and ¶ P < 0.001

Article Snippet: Assoc., Birmingham, AL, USA; rabbit anti-chicken IgY horseradish peroxidase conjugate was from Promega Corporation, Madison, WI, USA; mouse cytokine detecting ELISA kits were from Wuhan Boster Bio-Tech.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Acta Neuropathologica Communications

Article Title: Temporal tracking of microglial and monocyte single-cell transcriptomics in lethal flavivirus infection

doi: 10.1186/s40478-023-01547-4

Figure Lengend Snippet: Single-cell RNA sequencing analysis on resident and infiltrating myeloid cells from the virus-infected CNS. a Schematic showing scRNA-seq pipeline. Microglia from mock-infected and microglia and MCs from WNV-infected brains at dpi 5 and 7 were flow cytometrically sorted, barcoded and pooled in equal numbers for cell capture on the BD Rhapsody. Libraries were sequenced on the Illumina NextSeq 1000 and processed in SevenBridges and Seurat. b UMAP plot showing 6 microglia and 3 MC clusters. c, d Percent ( c ) and number ( d ) of each microglia and MC cluster out of total microglia and MC populations in mock-infected and infected brains at dpi 5 and 7. e UMAP plot coloured by cell type and timepoint and overlaid with sample identifiers/barcode numbers. f Dot plot showing the expression of nominal microglia and MC genes in total microglia and MC populations in mock-infected and infected brains at dpi 5 and 7. g Heatmap showing the top differentially expressed genes in total microglia and MC populations in mock-infected and infected brains at dpi 5 and 7. Data is presented from one independent experiment with four mice per group

Article Snippet: Cells were sorted into FBS and kept on ice until they were barcoded with the single-cell mouse multiplexing kit (BD Biosciences) which uses an anti-CD45 antibody.

Techniques: RNA Sequencing Assay, Infection, Expressing

Journal: Acta Neuropathologica Communications

Article Title: Temporal tracking of microglial and monocyte single-cell transcriptomics in lethal flavivirus infection

doi: 10.1186/s40478-023-01547-4

Figure Lengend Snippet: Microglia and MCs differentially promote CD4 + T cell IFN-γ production and CD8 + T cell proliferation. a Summary table showing changes in the frequency of IFN-γ + and BrdU + T cells with microglia and MC depletion in WNV-infected mice. b Flow cytometric dot plots showing the expression of IFN-γ in fully-stained samples and a fluorescence-minus-one (FMO, i.e. , without anti-IFN-γ) in non-stimulated T cells from mice fed control ( Ctrl ) or PLX5622-formulated chow ( PLX ) until sacrifice at dpi 7. c , d Number and frequency IFN-γ-producing T cells, geometric mean of IFN-γ in T cells and correlation analysis between the number of microglia and frequency of IFN-γ-producing T cells from single-cell brain suspensions from Ctrl and PLX mice sacrificed at dpi 5 ( c ) or dpi 7 ( d ) . e Number and frequency IFN-γ-producing T cells, geometric mean of IFN-γ in T cells and correlation analysis between the number of Ly6C hi MCs and frequency of IFN-γ-producing T cells from single-cell brain suspensions from anti-Ly6C-treated mice sacrificed at dpi 7 ( e ). f–i Frequency of BrdU + proliferating T cells in WNV-infected mice treated with PLX5622 and culled at dpi 5 ( f ) or 7 ( g ) or in WNV-infected mice treated with monoclonal antibodies to block monocyte infiltration and culled at dpi 5 ( h ) or 7 ( i ) . Data is presented as mean ± SEM from one or two independent experiments with 3–8 mice per group

Article Snippet: Cells were sorted into FBS and kept on ice until they were barcoded with the single-cell mouse multiplexing kit (BD Biosciences) which uses an anti-CD45 antibody.

Techniques: Infection, Expressing, Staining, Fluorescence, Blocking Assay